Cell-free protein expression using double-stranded concatemer DNA

摘要

A method for in vitro transcription and translation using double-stranded concatemer DNA in eukaryotic cell-free expression systems is provided. These methods include (a) contacting the double-stranded concatemer DNA with a eukaryotic cell-free expression system, and (b) expressing a protein from the double-stranded concatemer DNA in vitro in a eukaryotic cell-free expression system. Includes steps. The double-stranded concatemer DNA comprises multiple tandem repeat sequences. Multiple tandem repeat sequences include an expression sequence comprising a promoter, a cap-independent translation element (CITE), and an open reading frame. The final concentration of double-stranded concatemer DNA in a eukaryotic cell-free expression system ranges from about 0.1 ng / μL to about 35 ng / μL. RCA product DNA can be used as double stranded concatemer DNA for this method.